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[1]祖金珊,徐世文 陆信曜,宗 红,等.整合型1,2,4-丁三醇重组菌的构建及共底物发酵*[J].应用与环境生物学报,2019,25(04):1-9.[doi:Doi: 10.19675/j.cnki.1006-687x.2018.11006]
 ZU Jinshan,XU Shiwen,et al.Construction of an 1,2,4-butanetriol integrated strain and its co-substrate fermentation*[J].Chinese Journal of Applied & Environmental Biology,2019,25(04):1-9.[doi:Doi: 10.19675/j.cnki.1006-687x.2018.11006]
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整合型1,2,4-丁三醇重组菌的构建及共底物发酵*()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
25卷
期数:
2019年04期
页码:
1-9
栏目:
研究论文
出版日期:
2019-08-31

文章信息/Info

Title:
Construction of an 1,2,4-butanetriol integrated strain and its co-substrate fermentation*
文章编号:
201811006
作者:
祖金珊1 2 徐世文2 陆信曜1 2 宗 红1 2 诸葛斌1 2**
1江南大学糖化学与生物技术教育部重点实验室 无锡 214122 2江南大学工业生物技术教育部重点实验室,生物工程学院,工业微生物研究中心 无锡 214122
Author(s):
ZU Jinshan1 2 XU Shiwen2 LU Xinyao1 2 ZONG Hong1 2 & ZHUGE Bin1 2**
1 Key Laboratory of Carbohydrate Chemist ry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China 2Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Laboratory of Industrial Microorganisms, School of Biotechnology, Jiangnan University, Wuxi 214122, China
关键词:
124-丁三醇大肠杆菌磷酸转移酶系统混糖发酵整合表达
Keywords:
124-butanetriol Escherichia coli phosphotransferase system mixed-sugar fermentation integrated expression
DOI:
Doi: 10.19675/j.cnki.1006-687x.2018.11006
摘要:
1,2,4-丁三醇(BT)是重要的非天然化学品。为构建整合型BT合成菌株,实现木糖、葡萄糖共底物发酵,通过Red系统将基因kivD、xdh整合至Escherichia coli基因组的xylAB、ptsHI、ptsG、crr位点,并尝试利用廉价的乳糖替代IPTG诱导外源基因表达。结果表明,外源基因整合至xylAB后,生物量降低28%,重组菌E.coli W021能够代谢木糖合成BT(0.7 g/L)。添加葡萄糖为共底物后生物量提高36%,但碳分解代谢抑制作用限制了木糖的代谢,BT产量降低14%。进一步整合代谢基因至不同的磷酸转移酶系统(PTS)位点,其中整合至ptsHI基因后BT产量最高,达到2.8 g/L。优化葡萄糖、木糖浓度后,BT产量提高到3.6 g/L,进一步优化乳糖替代IPTG后BT产量为1.9 g/L。最后经发酵罐优化,BT产量提高到3.9 g/L,转化率为0.3 mol·mol-1。本研究构建整合型菌株在廉价乳糖诱导下共底物发酵合成BT,为后续放大研究提供了借鉴。
Abstract:
1,2,4-butanetriol (BT) is an important non-natural chemical. The exogenous genes kivD and xdh were integrated into xylAB site of Escherichia coli genome by R ed system to construct BT production integrated strain E.coli W021. To realize glucose/xylose co-fermentation, kivD and xdh were respectively integrated into ptsHI, ptsG, and crr sites in E.coli W021 to obtain recombinant strains E.coli W031, E.coli W032, and E.coli W033. N on-toxic and cheap lactose was used to replace IPTG for inducing expression of exogenous gene. The results showed the recombinant strain E.coli W021 produced 0.7 g/L BT from xylose, with a 28% decrease of biomass. After adding glucose as co-substrate, the biomass was increased by 36% . However, xylose metabolism was inhibited due to carbon catabolite repression (CCR), resulting in a BT production decrease of 1 4%. Modifying the phosphotransferase system (PTS) to alleviate the CCR effect, E.coli W031 produced BT at the highest titer of 2.8 g/L. BT production was further increased to 3.6 g/L by optimization of glucose and xylose concentrations. Additionally, s electing lactose instead of IPTG as inducer, the BT yield was 1.9 g/L. Finally, the BT yield reached 3.9 g/L and the conversion rate was 0.3 mol·mol-1 during batch fermentation . In this study, integrated strain was used to synthesize BT induced by lactose under co-substrate fermentation, which provided a reference for subsequent amplification studies.

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备注/Memo

备注/Memo:
收稿日期:2018-11-05 接受日期 Accepted: 2019-01-07?
*国家自然科学基金青年科学基金项目(21708016)、国家轻工技术与工程一流学科自主课题(LITE2018-01)资助?
**通讯作者(E-mail: bzhuge@126.com)
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更新日期/Last Update: 2019-01-24